Journal
ENDOCRINOLOGY
Volume 156, Issue 7, Pages 2492-2502Publisher
ENDOCRINE SOC
DOI: 10.1210/en.2014-1890
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Funding
- Swedish Research Council
- Swedish Foundation for Strategic Research
- Avtal om Lakarutbildning och Forskning research from Sahlgrenska University Hospital
- Lundberg Foundation
- Torsten and Ragnar Soderberg's Foundation
- Novo Nordisk Foundation
- Cancer Foundation Finland sr [140154, 130138] Funding Source: researchfish
- Novo Nordisk Fonden [NNF13OC0004839, NNF14OC0009883, NNF15OC0015902, NNF14OC0010513, NNF13OC0005785] Funding Source: researchfish
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Accurate measurement of sex steroid concentrations in rodent serum is essential to evaluate mouse and rat models for sex steroid-related disorders. The aim of the present study was to develop a sensitive and specific gas chromatography-tandem mass spectrometry (GC-MS/MS) method to assess a comprehensive sex steroid profile in rodent serum. A major effort was invested in reaching an exceptionally high sensitivity for measuring serum estradiol concentrations. We established a GC-MS/MS assay with a lower limit of detection for estradiol, estrone, T, DHT, progesterone, androstenedione, and dehydroepiandrosterone of 0.3, 0.5, 4.0, 1.6, 8, 4.0, and 50 pg/mL, respectively, whereas the corresponding values for the lower limit of quantification were 0.5, 0.5, 8, 2.5, 74, 12, and 400 pg/mL, respectively. Calibration curves were linear, intra-and interassay coefficients of variation were low, and accuracy was excellent for all analytes. The established assay was used to accurately measure a comprehensive sex steroid profile in female rats and mice according to estrous cycle phase. In addition, we characterized the impact of age, sex, gonadectomy, and estradiol treatment on serum concentrations of these sex hormones in mice. In conclusion, we have established a highly sensitive and specific GC-MS/MS method to assess a comprehensive sex steroid profile in rodent serum in a single run. This GC-MS/MS assay has, to the best of our knowledge, the best detectability reported for estradiol. Our method therefore represents an ideal tool to characterize sex steroid metabolism in a variety of sex steroid-related rodent models and in human samples with low estradiol levels.
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